GSM7658938: Mnase_bcl7ab_IP2; Arabidopsis thaliana; MNase-Seq
Lysates were clarified from sonicated protein-DNA complexes were isolated with GFP antibody. About 2 ng of ChIP DNA immuoprecipitated by anti-GFP antibody was used for ChIP-seq library construction. Blunting end, adapter ligation and amplification were carried out using Illumina Genomic DNA Sample Prep Kits according to the manufacturer's protocol. Novaseq was used for high-throughput sequencing of the ChIP-seq library.  The MNase assays were carried out as described previously85 with minor modifications. Approximately 0.4g of seedlings were used per sample. For MNase treatment, the prepared nuclei were resuspended in prewarmed MNase digestion buffer (20 mM Tris-HCl, pH 8.0, 5 mM NaCl, and 2.5 mM CaCl2), followed by the addition of 0.5 units of MNase (Takara, Cat. No. 2910A,) and incubation (10 min at 37◦C with periodic agitation). Finally, the mono-nucleosome DNA was isolated from 2% agarose gels and quantified by the Qubit dsDNA HS assay kit (ThermoFisher, Cat. No. Q32851).